小鼠细胞中8-氧代鸟嘌呤的转录偶联修复：ogg1蛋白是修复非转录序列所必需的，但不是转录序列所必需的。
Transcription coupled repair of 8-oxoguanine in murine cells: the ogg1 protein is required for repair in nontranscribed sequences but not in transcribed sequences.
作者信息
Le Page F, Klungland A, Barnes D E, Sarasin A, Boiteux S
机构信息
Laboratoire de Radiobiologie de l'ADN, UMR217, Commissariat à l'Energie Atomique-Centre National de la Recherche Scientifique, BP6, 92265-Fontenay aux Roses, France.
出版信息
Proc Natl Acad Sci U S A. 2000 Jul 18;97(15):8397-402. doi: 10.1073/pnas.140137297.
Abstract
To assess the role of the Ogg1 DNA glycosylase in the transcription-coupled repair (TCR) of the mutagenic lesion, 7, 8-dihydro-8oxoguanine (8-OxoG), we have investigated the removal of this lesion in wild-type and ogg1(-/-) null mouse embryo fibroblast (MEF) cell lines. We used nonreplicating plasmids containing a single 8-OxoG.C base pair in a different assay that allowed us to study the removal of 8-OxoG located in a transcribed sequence (TS) or in a nontranscribed sequence (NTS). The results show that the removal of 8-OxoG in a wild-type MEF cell line is faster in the TS than in the NTS, indicating TCR of 8-OxoG in murine cells. In the homozygous ogg1(-/-) MEF cell line, 8-OxoG was not removed from the NTS whereas there was still efficient 8-OxoG repair in the TS. Expression of the mouse Ogg1 protein in the homozygous ogg1(-/-) cell line restored the ability to remove 8-OxoG in the NTS. Therefore, we have demonstrated that Ogg1 is essential for the repair of 8-OxoG in the NTS but is not required in the TS. These results indicate the existence of an Ogg1-independent pathway for the TCR of 8-OxoG in vivo.
摘要
为了评估Ogg1 DNA糖基化酶在诱变损伤7,8 - 二氢 - 8 - 氧代鸟嘌呤（8 - OxoG）的转录偶联修复（TCR）中的作用，我们研究了野生型和ogg1(-/-)基因敲除小鼠胚胎成纤维细胞（MEF）系中该损伤的去除情况。我们在不同的实验中使用了含有单个8 - OxoG.C碱基对的非复制性质粒，这使我们能够研究位于转录序列（TS）或非转录序列（NTS）中的8 - OxoG的去除情况。结果表明，野生型MEF细胞系中TS区域的8 - OxoG去除速度比NTS区域快，这表明小鼠细胞中存在8 - OxoG的TCR。在纯合ogg1(-/-) MEF细胞系中，NTS区域的8 - OxoG没有被去除，而TS区域仍有高效的8 - OxoG修复。在纯合ogg1(-/-)细胞系中表达小鼠Ogg1蛋白恢复了去除NTS区域8 - OxoG的能力。因此，我们证明了Ogg1对于NTS区域8 - OxoG的修复至关重要，但在TS区域则不是必需的。这些结果表明体内存在一条不依赖Ogg1的8 - OxoG的TCR途径。
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